Gel express: a novel frugal method quantifies gene relative expression in conventional RT-PCR

Background Real-time PCR system is a valuable scientific mainstream needed for quantifying specific gene expression. Nevertheless, compared with conventional PCR, the real-time PCR system is extremely expensive and not affordable for limited or mid-budget research laboratories. Here, a novel, doable and low-cost recipe (referred to as gel express) is developed to quantify gene expression using conventional RT-PCR assay. The novelty of the gel express method is based on replacing crossing point (CP) values with integrated density (IntDen) values of PCR amplicon bands in real-time PCR regular mathematical formulas. Results In this work, gene expression profiles of two different rice stress-marker genes (OsCYP94C2a and OsLOX8) were quantified in response to mechanical wounding at different time points (0, 30, 60, and 150 min). In the gel express method, the free software ImageJ was employed to measure integrated density (IntDen) values of PCR amplicon bands in agarose gel images. IntDen values were then used instead of crossing point (CP) values according to the following modified formula: [EIntDen(ref)/EIntDen(target)]sample ÷ [EIntDen(ref)/EIntDen(target)]control. Gene relative expression profiles (dynamic expression pattern) quantified by gel express method in both genes were highly comparable with real-time RT-PCR. R2 values were 0.9976 and 0.9975 in OsCYP94C2a and OsLOX, respectively. PCR amplification efficiency (E) for all studied genes could be calculated depending on IntDen values through experimentally designed calibration curves. PCR amplification efficiencies with all studied genes obtained by gel express were all in the accepted range. For better-visualized PCR amplicons thus detectable biological effects between treatments, the number of PCR cycles applied in gel express method (IntCyc) was experimentally estimated to be 29 cycles. Conclusions Gel express is a novel, cost-effective and feasible recipe for quantifying gene relative expression in conventional RT-PCR. The expression pattern quantified by gel express is highly comparable and fits the expression data revealed by the used real-time PCR system.

Manipulation of Time- and Frequency-Domain Dynamics by Magnon-Magnon Coupling in Synthetic Antiferromagnets

Magnons (the quanta of spin waves) could be used to encode information in beyond Moore computing applications. In this study, the magnon coupling between acoustic mode and optic mode in synthetic antiferromagnets (SAFs) is investigated by micromagnetic simulations. For a symmetrical SAF system, the time-evolution magnetizations of the two ferromagnetic layers oscillate in-phase at the acoustic mode and out-of-phase at the optic mode, showing an obvious crossing point in their antiferromagnetic resonance spectra. However, the symmetry breaking in an asymmetrical SAF system by the thickness difference, can induce an anti-crossing gap between the two frequency branches of resonance modes and thereby a strong magnon-magnon coupling appears between the resonance modes. The magnon coupling induced a hybridized resonance mode and its phase difference varies with the coupling strength. The maximum coupling occurs at the bias magnetic field at which the two ferromagnetic layers oscillate with a 90° phase difference. Besides, we show how the resonance modes in SAFs change from the in-phase state to the out-of-phase state by slightly tuning the magnon-magnon coupling strength. Our work provides a clear physical picture for the understanding of magnon-magnon coupling in a SAF system and may provide an opportunity to handle the magnon interaction in synthetic antiferromagnetic spintronics.

In Situ Pinpoint Photopolymerization of Phos-Tag Polyacrylamide Gel in Poly(dimethylsiloxane)/Glass Microchip for Specific Entrapment, Derivatization, and Separation of Phosphorylated Compounds

An improved method for the online preconcentration, derivatization, and separation of phosphorylated compounds was developed based on the affinity of a Phos-tag acrylamide gel formed at the intersection of a polydimethylsiloxane/glass multichannel microfluidic chip toward these compounds. The acrylamide solution comprised Phos-tag acrylamide, acrylamide, and N,N-methylene-bis-acrylamide, while 2,2′-azobis[2-methyl-N-(2-hydroxyethyl)propionamide] was used as a photocatalytic initiator. The Phos-tag acrylamide gel was formed around the channel crossing point via irradiation with a 365 nm LED laser. The phosphorylated peptides were specifically concentrated in the Phos-tag acrylamide gel by applying a voltage across the gel plug. After entrapment of the phosphorylated compounds in the Phos-tag acrylamide gel, 5-(4,6-dichlorotriazinyl)aminofluorescein (DTAF) was introduced to the gel for online derivatization of the concentrated phosphorylated compounds. The online derivatized DTAF-labeled phosphorylated compounds were eluted by delivering a complex of phosphate ions and ethylenediamine tetraacetic acid as the separation buffer. This method enabled sensitive analysis of the phosphorylated peptides.

Excellent Ultracold Molecular Candidates From Group VA Hydrides: Whether Do Nearby Electronic States Interfere?

By means of highly accurate ab initio calculations, we identify two excellent ultracold molecular candidates from group VA hydrides. We find that NH and PH are suitable for the production of ultracold molecules, and the feasibility and advantage of two laser cooling schemes are demonstrated, which involve different spin-orbit states (A3Π2 and X3Σ1− ). The internally contracted multireference configuration interaction method is applied in calculations of the six low-lying Λ-S states of NH and PH with the spin-orbit coupling effects included, and excellent agreement is achieved between the computed and experimental spectroscopic data. We find that the locations of crossing point between the A3Π and Σ−5 states of NH and PH are higher than the corresponding v′ = 2 vibrational levels of the A3Π state indicating that the crossings with higher electronic states would not affect laser cooling. Meanwhile, the extremely small vibrational branching loss ratios of the A3Π2 → a1Δ2 transition for NH and PH (NH: 1.81 × 10–8; PH: 1.08 × 10–6) indicate that the a1Δ2 intermediate electronic state will not interfere with the laser cooling. Consequently, we construct feasible laser-cooling schemes for NH and PH using three lasers based on the A3Π2 → X3Σ1− transition, which feature highly diagonal vibrational branching ratio R00 (NH: 0.9952; PH: 0.9977), the large number of scattered photons (NH: 1.04×105; PH: 8.32×106) and very short radiative lifetimes (NH: 474 ns; PH: 526 ns). Our work suggests that feasible laser-cooling schemes could be established for a molecular system with extra electronic states close to those chosen for laser-cooling.

On-line Measurement Method and Technical Scheme of Transmission Line Crossing Point Based on Machine Vision

With the increasing demand for high voltage and UHV transmission, higher and higher requirements are put forward for transmission line monitoring and fault diagnosis. The traditional line measurement method has many shortcomings, which cannot ensure the measurement accuracy, measurement efficiency and high cost. Based on this, this paper first studies the principle of machine vision on-line measurement, then analyses the on-line measurement process of transmission line crossing point based on machine vision, and finally gives the error source and amelioration measures of transmission line crossing point on-line measurement.

Turning Wages into Capital Differentiation on the Market for Unsecured Loans in the United States, 1900-1945

In this article, we show how interpretive battles about compliance can lead to regulatory differentiation and, in turn, market segmentation. To do so, we study the evolution of unsecured lending in the United States, between 1900 and 1945. In the early 20th century, a large segment of the workforce relied on their wages to access credit: this required the “legal coding” of labor income into capital, where lenders would offer advances in exchange for a lien over future revenues. Regulating these transactions raised conflicts between Progressive reformers, lenders and, after 1929, federal regulators, which spanned over five decades. The historical comparison of three states—Illinois, New York and Georgia—, shows that local discussions revolved around three outcomes—legal status, pricing method and collateralization—, the issue of which led to distinct regulatory paths and market configurations at the state level. Finally, the New Deal policies created an additional strand of federal coding, furthering market divides between unregulated payday lenders, non-bank credit companies, and commercial banks. On financial markets, discussions about compliance often revolve around calculative technologies, and we suggest this as a possible crossing point between STS analyses of capitalization devices and Pistor’s theory of capital modulation.

Zero-Crossing Point Detection of Sinusoidal Signal in Presence of Noise and Harmonics Using Deep Neural Networks

Zero-crossing point detection is necessary to establish a consistent performance in various power system applications, such as grid synchronization, power conversion and switch-gear protection. In this paper, zero-crossing points of a sinusoidal signal are detected using deep neural networks. In order to train and evaluate the deep neural network model, new datasets for sinusoidal signals having noise levels from 5% to 50% and harmonic distortion from 10% to 50% are developed. This complete study is implemented in Google Colab using deep learning framework Keras. Results shows that the proposed deep learning model is able to detect zero-crossing points in a distorted sinusoidal signal with good accuracy.

Editor's Note

Charles Bernstein is a major crossing point for poetry and poetics. His art and discussion of poetry exploit and develop the vernaculars of language as they echo across time and international borders and national languages.

Looking for the Fusion of Cultural Environment and Modernity in a Cityscape: Western Gate in Jelgava

The research is based on the examination of the cityscape transformation processes and a search for the fusion of the cultural and historical space and the trends of contemporary modernism in architecture. Over the last three centuries, Jelgava (Mitau), the capital of the former Dutchy of Courland, has changed the height, the density and the structure of its building. The process of transformation was determined by crisis situations in the state, by rapid growth of the trade and domestic economy, as well as by the period of state independence and downfall of the national economy. Splendour and misery of the city has raised and destroyed houses, parks and churches in Jelgava.The historical map of the city originates in the beginning of the 18th century on the left bank of the Lielupe River with a linear building canvas formed by small wooden houses and a net of dirt roads. During the 19th-20th centuries, the city is developing radially around the ancient central built-up area, sketching the structure of the city map based on the network of the trading routes: the Western gate – Dobele, the Northern gate – sea, the Southern gate – Lithuania. The East is a crossing point to reach Jelgava Palace. The direction to Riga led along the river, as the eastern part was a marshland.